A Bacillus amyloliquefaciens with dual functions of degrading inorganic phosphorus and preventing diseases
A technology of amylolytic spores and inorganic phosphorus, applied in the field of agricultural microorganisms, can solve the problem of single function of bacillus, achieve the effects of less resistance to drugs, improve utilization rate, and reduce the number of spraying
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Embodiment 1
[0048]Embodiment 1 Separation screening process and classification identification of a fracture in the fracture plant WKPHO-12
[0049](1) Separation screening process of the strain of the present invention:
[0050]In September 2015, five points were collected at five o'clock in the Alum hill phosphorus mine in Zhangjiakou City, Hebei Province, 200g per share. Siders 1.0g of wind-dried soil samples were added to the triangular bottle with sterilized glass beads, and then 99 ml of sterile water, allowed to stand for 20 min, 30 ° C on the shaker, 180R / min thoroughly oscillated for 30 min, then according to the 10-fold dilution method Dilution of gradient, take 10 separately-310-410-5100 μL of the dilution, coated on an inorganic phosphorus medium, each concentration of 3 repetitions; after painting, stand in the clean station, in the absence of a bacteria, at 35 ° C constant temperature culture 5 ~ 7D. Through the transparent ring method, the molybdenum resistance is screened with a stra...
Embodiment 2
[0059]Example 2 Preparation of Microbacterial Agent containing WKPHO-12 strains
[0060]According to the following steps:
[0061](1) Activation of strains: Clear strain WKPHO-12 (which is prepared as CGMCC No. 14954) preserved at -80 ° C is activated (30 ° C), and the single colony is picked up in the LB tablet medium. The LB ramp medium was cultured at 30 ° C for 12 hours to activate a strain; the composition ingredients of the LB plate medium or Lb ramp medium and its weight ratio were: trypsin 10g, yeast extract 5 g, sodium chloride 5g, Agar powder 15g, water 1000ml;
[0062](2) Preparation of Seed Liquid: Loading LB liquid medium in a 250 mL triangular bottle (its composition ingredients and weight ratios are: trypsin 10g, yeast extract 5g, sodium chloride 5g, water 1000ml) 100ml, high pressure moist heat After sterilization, after the temperature is lowered to room temperature, the activated strain in which the activated strain is entered into a circulating ring step (1), and oscillati...
Embodiment 3
[0066]Example 3 Test of the determination of a fracture inorganic phosphorus capabilities of this fracture
[0067]Perform as follows:
[0068]The use of sterilization to activate a good WKPHO-12 strain in Example 2 (1) in the solution in which the inorganic phosphor plate medium (its composition ingredients and weight ratios are: glucose 10.0 g, (NH4)2SO40.5g, MGSO4· 7h2O 0.5g, NaCl 0.2g, CA3(Po42 5.0g, KCl 0.2g, MNSO4 0.03g, FESO4 0.003 g, agar 20.0 g, distilled water 1000 mL, pH: 7.0-8.0), and then placed at a constant temperature incubator in a 30 ° C for 5 days, and the diameter of the transparent loop and colonies were measured.
[0069]Results The fracture of this invention WKPHO-12 is containing CA3(Po4)2The inorganic phosphor plate medium produces a transparent circle having a diameter of 13.6 mm, indicating that the WKPHO-12 strain of the present invention can degrade inorganic phosphorus CA.3(Po4)2, With the potential of inorganic phosphorus in the soil.
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